Pepper variety nun 70064 pph

ABSTRACT

A new and distinct pepper variety NUN 70064 PPH is disclosed, as well as seeds and plants and heads or leaves thereof.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority of the U.S. Provisional Application No.63/343,868 filed on May 19, 2022, which is hereby incorporated byreference in its entirety.

FIELD OF DISCLOSURE

The disclosure relates to the field of plant breeding and, morespecifically, to the pepper variety NUN 70064 PPH. The disclosurefurther relates to vegetative reproductions of pepper variety NUN 70064PPH, methods for tissue culture of pepper variety NUN 70064 PPH, andmethods for regenerating a plant from such a tissue culture and also tophenotypic variants of pepper variety NUN 70064 PPH. The disclosure alsorelates to progeny of pepper variety NUN 70064 PPH and the hybridvarieties obtained by crossing pepper variety NUN 70064 PPH as a parentline with plants of other varieties or parents lines.

BACKGROUND OF THE DISCLOSURE

The goal of plant breeding is to combine various desirable traits in asingle variety. Such desirable traits may include greater yield,resistance to diseases, insects or other pests, tolerance to heat anddrought, better agronomic quality, higher nutritional value, enhancedgrowth rate, and improved fruit properties.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same genotype. A plant cross-pollinates if pollen comes to it from aflower of a different genotype. Plants that have been self-pollinatedand selected for (uniform) type over many generations become homozygousat almost all gene loci and produce a uniform population of truebreeding progeny of homozygous plants. A cross between two suchhomozygous plants of different lines produces a uniform population ofhybrid plants that are heterozygous for many gene loci. The extent ofheterozygosity in the hybrid is a function of the genetic distancebetween the parents. Conversely, a cross of two plants each heterozygousat a number of loci produces a segregating population of hybrid plantsthat differ genetically and are not uniform. The resultingnon-uniformity makes performance unpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants to makehybrids, and the evaluation of the hybrids resulting from the crosses.Pedigree breeding and recurrent selection are examples of breedingmethods that have been used to develop inbred plants from breedingpopulations. Those breeding methods combine the genetic backgrounds fromtwo or more plants or various other broad-based sources into breedingpools from which new lines are developed by selfing and selection ofdesired phenotypes. The new plants are evaluated to determine which havecommercial potential.

One crop species that has been subject to such breeding programs and isof particular value is the pepper. Pepper (Capsicum spp.) is naturally adiploid and the basic chromosome number of the genus is x=12, most are2n=2×=24, including the cultivated ones. A few wild species have 2n=26.Ploidy changes (both tetraploidy and haploidy) are relatively easy toinduce in Capsicum species. Doubled haploids have proved particularlyvaluable in the analysis of the genetically complex basis of someresistances to pests and diseases.

The genus Capsicum originated in American tropics. The fruit of mostspecies of Capsicum produce a strong burning sensation (pungency orspiciness) in the mouth of the unaccustomed eater due to the presence ofcapsaicin (methyl vanillyl nonanamide), a lipophilic chemical, making itas an important spice commodity. Capsaicin can be present in largequantities in the placental tissue (which holds the seeds), the internalmembranes, and to a lesser extent, the other fleshy parts of the fruitsof plants in this genus. The seeds themselves do not produce anycapsaicin. The amount of capsaicin in the fruit is highly variable anddependent on genetics and environment, giving almost all types ofCapsicum varied amounts of perceived heat.

The most recognizable Capsicum without capsaicin is the bell pepper, acultivar of Capsicum annuum, which has a zero rating on the Scovillescale. The lack of capsaicin in bell peppers is due to a recessive genethat eliminates capsaicin and, consequently, the “hot” taste usuallyassociated with the rest of the Capsicum family.

Pepper can be classified according to its target market: fresh marketand processing peppers. Peppers for the fresh market require that thefruits are firm, shiny and have fresh green calyx and stem. They aretypically consumed fresh as a snack or used in salad or sandwiches or asa cooked vegetable. On the other hand, processing peppers are used forfreezing or dehydrating and can be dried, ground as spices andprocessed, e.g., pickled, canned, brined or in salsas.

In the United States, majority of the peppers produced is the bellpepper (i.e., sweet pepper), which are mainly marketed fresh. Californiaand Florida are the lead producers of bell peppers. Bell peppers areavailable year-round with supply at the greatest volume from May to Julyand March to April in California and Florida, respectively. Bulk of thebell peppers grown and harvested are green, but premium is given tocolored bell peppers (i.e., red, yellow).

Both hybrids and open-pollinated varieties are used for production inthe United States, with a growing trend in the use of seeded hybridvarieties. Hybrid varieties offer the advantages of easy combination ofdominant and recessive traits, such as disease resistance, from a set ofinbred parents, as well as careful control of parentage.

Advances in biotechnology have also resulted in genetically engineeredpepper plants with improved traits. For example, fungal resistant pepperplants comprising a PepEST or PepDef gene where the expression of thenucleic acid sequence in the plant resulted in increased resistance tofungal infection, see, e.g., U.S. Pub. No. 2006/0037100, which herebyincorporated by reference in its entirety.

While breeding efforts to date have provided a number of useful peppervarieties with beneficial traits, there remains a great need in the artfor new varieties with further improved traits. Such plants wouldbenefit farmers and consumers alike by improving crop yields and/orquality. Breeding objectives include resistance to pests and diseases,improvement of fruit quality, protection against biotic and abioticstresses, varying the color, texture and flavor of the fruit, optimizingflesh thickness, yield, suitability to various climatic circumstances,heat, solid content (% dry matter), or sugar content.

SUMMARY OF VARIOUS ASPECTS OF THE DISCLOSURE

The disclosure provides for a pepper variety NUN 70064 PPH, productsthereof, and methods of using the same. NUN 70064 PPH is a jalapenopepper variety for the fresh market and is suitable for growing in theopen field.

The disclosure also provides a pepper plant, or part thereof, having allof the physiological and morphological characteristics of pepper varietyNUN 70064 PPH when grown under the same environmental conditions.

In another aspect, pepper variety NUN 70064 PPH or a progeny thereofcomprises resistance to Tomato Spotted Wilt Virus (TSWV), measuredaccording to UPOV standards described in TG/76/8.

The disclosure also provides for a progeny of pepper variety NUN 70064PPH. In another aspect, the disclosure provides a plant or a progenyretaining all or all but one, two, or three of the “distinguishingcharacteristics” or all or all but one, two, or three of the“morphological and physiological characteristics” of pepper variety NUN70064 PPH and methods for producing that plant or progeny.

In one aspect, the disclosure provides a plant or a progeny having allthe physiological and morphological characteristics of variety NUN 70064PPH when grown under the same environmental conditions. In anotheraspect, the plant or progeny has all or all but one, two, or three ofthe physiological and morphological characteristics of pepper varietyNUN 70064 PPH when measured under the same environmental conditions ande.g., evaluated at significance levels of 1%, 5% or 10% significance(which can also be expressed as a p-value) for quantitativecharacteristics and determined by type or degree for non-quantitativecharacteristics, wherein a representative sample of seed of peppervariety NUN 70064 PPH has been deposited under Accession Number NCIMB______. In another aspect, the plant or progeny has all or all but one,two or three of the physiological and morphological characteristics aslisted in Table 1 for pepper variety NUN 70064 PPH when measured underthe same environmental conditions and e.g., evaluated at significancelevels of 1%, 5% or 10% significance (which can also be expressed as ap-value) for quantitative characteristics and determined by type ordegree for non-quantitative characteristics.

In one aspect, the disclosure provides, a seed of pepper variety NUN70064 PPH, wherein a representative sample of said seed has beendeposited under Accession Number NCIMB ______. The disclosure alsoprovides for a plurality of seeds of pepper variety NUN 70064 PPH. Theseed of pepper variety NUN 70064 PPH may be provided as an essentiallyhomogeneous population of pepper seed. The population of seed of peppervariety NUN 70064 PPH may be particularly defined as being essentiallyfree from other seed. The seed population may be grown into plants toprovide an essentially homogeneous population of pepper plants asdescribed herein.

The disclosure also provides a plant grown from a seed of pepper varietyNUN 70064 PPH and a plant part thereof.

The disclosure furthermore provides a pepper fruit produced on a plantgrown from a seed of pepper variety NUN 70064 PPH.

In another aspect, the disclosure provides, a seed growing or grown on aplant of variety NUN 70064 PPH (i.e., produced after pollination of theflower of pepper variety NUN 70064 PPH).

In another aspect, the disclosure provides for a hybrid variety ofpepper called NUN 70064 PPH.

In another aspect, the disclosure provides for a plant part obtainedfrom variety NUN 70064 PPH, wherein said plant part is: a fruit, aharvested fruit, a part of a fruit, a leaf, a part of a leaf, pollen, anovule, a cell, a petiole, a shoot or a part thereof, a stem or a partthereof, a root or a part thereof, a root tip, a cutting, a seed, a partof a seed, seed coat or another maternal tissue which is part of a seedgrown on said variety, a hypocotyl, a cotyledon, a scion, a stock, arootstock, a pistil, an anther, or a flower or a part thereof. Fruitsare particularly important plant parts. In a further aspect, the plantpart obtained from variety NUN 70064 PPH is a cell, optionally a cell ina cell or tissue culture. That cell may be grown into a plant of varietyNUN 70064 PPH.

The disclosure also provides a cell culture of pepper variety NUN 70064PPH, and a plant regenerated from pepper variety NUN 70064 PPH, whereinsaid plant has all the characteristics of pepper variety NUN 70064 PPHwhen grown under the same environmental conditions, as well as methodsfor culturing and regenerating pepper variety NUN 70064 PPH.Alternatively, a regenerated plant may have one characteristic that isdifferent from pepper variety NUN 70064 PPH, and which otherwise has allof the physiological and morphological characteristics of pepper varietyNUN 70064 PPH.

The disclosure further provides a vegetatively propagated plant ofvariety NUN 70064 PPH having all or all but one, two, or three of themorphological and physiological characteristics pepper variety NUN 70064PPH when grown under the same environmental conditions.

In another aspect, the disclosure provides a method of producing apepper plant comprising crossing pepper variety NUN 70064 PPH withitself or with another pepper variety and selecting a progeny pepperplant from said crossing.

The disclosure also provides a method of producing a pepper plantderived from pepper variety NUN 70064 PPH.

In a further aspect, the disclosure provides a method of producing ahybrid pepper seed comprising crossing a first parent pepper plant witha second parent pepper plant and harvesting the resultant hybrid pepperseed, wherein said first pepper plant or second pepper plant is peppervariety NUN 70064 PPH. Also provided is a hybrid pepper plant producedfrom crossing a first pepper plant and second pepper plant andharvesting the resultant hybrid pepper seed, wherein said first pepperplant or second pepper plant is pepper variety NUN 70064 PPH. Moreover,the hybrid pepper plant grown from the hybrid pepper seed is provided.

In another aspect, the disclosure provides a method of introducing asingle locus conversion into the plant of variety NUN 70064 PPH, whereina representative sample of seed of said pepper variety has beendeposited under Accession Number NCIMB ______, wherein the plantcomprises the single locus conversion and otherwise has all of thephysiological and morphological characteristics of the plant of peppervariety NUN 70064 PPH.

In yet another aspect, the disclosure provides a method for introducinga desired trait into pepper variety NUN 70064 PPH, said method comprisestransforming the plant of pepper variety NUN 70064 PPH with a transgenethat confers the desired trait, wherein the transformed plant containsthe desired trait and otherwise has all of the physiological andmorphological characteristics of the plant of pepper variety NUN 70064PPH.

The disclosure also provides a method of producing a modified pepperplant with a desired trait, wherein the method comprises mutating atarget gene by targeted gene editing in a pepper plant or plant part ofpepper variety NUN 70064 PPH, wherein the target gene modifies a desiredtrait.

In one aspect, the single locus conversion or desired trait is yield,storage properties, color, flavor, size, firmness, enhanced nutritionalquality, post-harvest quality, male sterility, herbicide tolerance,insect resistance, pest resistance, disease resistance, environmentalstress tolerance, modified carbohydrate metabolism, modified proteinmetabolism, or ripening.

In another aspect, the disclosure provides a container comprising theplant, plant part, or seed of pepper variety NUN 70064 PPH.

Also provided is a food, a feed, or a processed product comprising aplant part of pepper variety NUN 70064 PPH, wherein the plant part is afruit or a part thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the immature fruit of pepper variety NUN 70064 PPH.

DEFINITIONS

“Pepper” refers herein to plants of the species Capsicum annuum orfrutescens, and fruits thereof. The most commonly eaten part of a pepperis the fruit. The fruit comprises a stem or peduncle or pedicel, calyx,placenta, fruit wall, veins, shoulder, base, apex, locule or lobe,septa, exocarp, mesocarp, endocarp, pericarp, optionally secondarypeppers, optionally capsaicin glands and optionally seed. The stem orpeduncle or pedicel, calyx, placenta, fruit wall, veins, shoulder, base,apex, locule or lobe, septa, exocarp, mesocarp, endocarp, pericarp,secondary peppers, capsaicin glands and seed coat of the seed arematernal tissues, and thus they are genetically identical to the planton which they grow.

“Cultivated pepper” refers to plants of Capsicum annuum, or a closelyrelated species, e.g., varieties, breeding lines or cultivars of thespecies C. annuum as well as crossbreds thereof, or crossbreds withother Capsicum species, cultivated by humans and having good agronomiccharacteristics.

The terms “pepper plant designated NUN 70064 PPH,” “NUN 70064 PPH,” “NUN70064,” “NUN 70064 F1,” “70064 PPH,” or “pepper 70064,” are usedinterchangeably herein and refer to a pepper plant of variety NUN 70064PPH, representative seed of which has been deposited under AccessionNumber NCIMB ______.

A “seed of NUN 70064 PPH” refers to a pepper seed which can be growninto a plant of variety NUN 70064 PPH, wherein a representative sampleof viable seed of said pepper variety has been deposited under AccessionNumber NCIMB ______. A seed can be in any stage of maturity, forexample, a mature, viable seed, or an immature, non-viable seed. A seedcomprises an embryo and maternal tissues.

An “embryo of NUN 70064 PPH” refers to an “F1 hybrid embryo” as presentin a seed of pepper variety NUN 70064 PPH, a representative sample ofsaid seed of said pepper variety has been deposited under AccessionNumber NCIMB ______.

A “seed grown on NUN 70064 PPH” refers to a seed grown on a mature plantof variety NUN 70064 PPH or inside a fruit of pepper variety NUN 70064PPH. The “seed grown on NUN 70064 PPH” contains tissues and DNA of thematernal parent, pepper variety NUN 70064 PPH. The “seed grown on NUN70064 PPH” contains an F2 embryo. When said seed is planted, it growsinto a first generation progeny plant of variety NUN 70064 PPH.

A “fruit of NUN 70064 PPH” refers to a fruit containing maternal tissuesof pepper variety NUN 70064 PPH as deposited under Accession NumberNCIMB ______. In one aspect, the fruit does not contain seed, i.e., thefruit is parthenocarpic. The skilled person is familiar with methods forinducing parthenocarpy. Those methods comprise chemically or geneticallyinducing parthenocarpy. Compounds suitable for chemically inducingparthenocarpy include auxins, gibberellins and cytokinins. Geneticparthenocarpy can be induced by CaARF8 mutants (see, e.g., Tiwari etal., BMC Plant Biology 2011, 11:143 DOI: 10.1186/1471-2229-11-143 orU.S. Pat. No. 8,492,619, which are herein incorporated by reference intheir entireties). A fruit can be in any stage of maturity, for example,comprising viable seed or comprising immature non-viable seed.

An “essentially homogeneous population of pepper seed” is a populationof seeds where at least 97%, 98%, 99% or more of the total population ofseed are seed of pepper variety NUN 70064 PPH.

An “essentially homogeneous population of pepper plants” is a populationof plants where at least 97%, 98%, 99% or more of the total populationof plants are plants of variety NUN 70064 PPH.

The phrase “essentially free from other seed” refers to a population ofseed where less than 3%, 2%, 1% or less of the total population of seedis seed that is not a pepper seed or, in another option, less than 3%,2%, 1% or less of the total population of seed is seed that is not seedof pepper variety NUN 70064 PPH.

“USDA descriptors” are the plant variety descriptors for pepper(Capsicum spp.) as published by the US Department of Agriculture,Agricultural Marketing Service, Plant Variety Protection Office,Beltsville, MD 20705, and which can be downloaded from the world wideweb at ams.usda.gov under services/plant-variety-protection/pvpo-c-formsunder pepper. “Non-USDA descriptors” are other descriptors suitable fordescribing pepper.

“UPOV descriptors” are the plant variety descriptors described forpepper in the “Guidelines for the Conduct of Tests for Distinctness,Uniformity and Stability, TG/76/8 (Geneva 2006, revised 2018), aspublished by UPOV (International Union for the Protection of NewVarieties and Plants, and which can be downloaded from the world wideweb at upov.int under edocs/tgdocs/en/tg076.pdf and is hereinincorporated by reference in its entirety. Likewise, “UPOV methods” todetermine specific parameters for the characterization of pepper aredescribed at upov.int.

“Calibration book Capsicum annuum L.” refers to the calibration book forpepper which provides guidance for describing a pepper variety, aspublished by Naktuinbow (December 2010, version 1) and based on the UPOVGuideline TG/76/8 and CPVO (Community Plant Variety Office) ProtocolCPVO-TP/076/2.

“RHS” refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Horticulture Society Enterprise Ltd RHS Garden;Wisley, Woking; Surrey GU236QB, UK, e.g., the RHS color chart: 2007.

“Reference Variety for NUN 70064 PPH” refers herein to variety Mixteco,a commercial variety from HM Clause, which has been planted in a trialtogether with pepper variety NUN 70064 PPH.

“Plant” includes the whole plant or any part or derivatives thereof,preferably having the same genetic makeup as the plant from which it isobtained.

“Plant part” includes any part of a plant, such as a plant organ (e.g.,harvested or non-harvested fruits), a plant cell, a plant protoplast, aplant cell tissue culture or a tissue culture from which a whole plantcan be regenerated, a plant cell that is intact in a plant, a clone, amicropropagation, plant callus, a plant cell clump, a plant transplant,a vegetative propagation, a seedling, a fruit, a harvested fruit, a partof a fruit, a leaf, a part of a leaf, pollen, an ovule, an embryo, apetiole, a shoot or a part thereof, a stem or a part thereof, a root ora part thereof, a root tip, a cutting, a seed, a part of a seed,hypocotyl, cotyledon, a scion, a graft, a stock, a rootstock, a pistil,an anther, and a flower or part thereof. Seed can be mature or immature.Pollen or ovules may be viable or non-viable. Also, any developmentalstage is included, such as seedlings, cuttings prior or after rooting,mature plants or leaves. Alternatively, a plant part may also include aplant seed which comprises the maternal tissues of pepper variety NUN70064 PPH and an embryo having one or two sets of chromosomes derivedfrom the parent plant, e.g., from pepper variety NUN 70064 PPH. Such anembryo comprises two sets of chromosomes derived from pepper variety NUN70064 PPH, if it is produced from self-pollination, while an embryoderived from cross-fertilization of pepper variety NUN 70064 PPH willcomprise only set of chromosomes from said variety.

“Rootstock” or “stock” refers to the plant selected for its roots, inparticular for the resistance of the roots to diseases or stress (e.g.,heat, cold, salinity etc.). Normally, the quality of the fruit of theplant providing the rootstock is less important.

“Scion” refers to a part of the plant attached to the rootstock. Thisplant is selected for its stems, leaves, flowers, or fruits. The scioncontains the desired genes to be duplicated in future production by thestock/scion plant and may produce the desired pepper fruit.

“Stock/scion” or grafted plant refers to a pepper plant comprising arootstock from one plant grafted to a scion from another plant.

“Harvest maturity” is referred to as the stage at which a pepper fruitis ripe or ready for harvest or the optimal time to harvest the fruitfor the market, for processing or for consumption. In one aspect,harvest maturity is the stage which allows proper completion of thenormal ripening.

“Harvested plant material” refers herein to plant parts (e.g., fruitsdetached from the whole plant), which have been collected for furtherstorage and/or further use.

“Yield” means the total weight of all pepper fruits harvested perhectare of a particular line or variety. It is understood that “yield”expressed as weight of all pepper fruits harvested per hectare can beobtained by multiplying the number of plants per hectare times the“yield per plant”.

“Marketable yield” means the total weight of all marketable pepperfruits, especially fruit that is not cracked, damaged or diseased,harvested per hectare of a particular line or variety. A “marketablefruit” is a fruit that has commercial value.

“Flavor” refers to the sensory impression of a food or other substance,especially a pepper fruit or fruit part (fruit flesh) and is determinedmainly by the chemical senses of taste and smell. Flavor is influencedby texture properties and by volatile and/or non-volatile chemicalcomponents (organic acids, lipids, carbohydrates, salts, etc.).

A plant having “all the physiological and morphological characteristics”of a referred-to-plant means a plant showing the physiological andmorphological characteristics of the referred-to-plant when grown underthe same environmental conditions, preferably in the same experiment;the referred-to-plant can be a plant from which it was derived, e.g.,the progenitor plant, the parent, the recurrent parent, the plant usedfor tissue- or cell culture, etc. A physiological or morphologicalcharacteristic can be a numerical characteristic or a non-numericalcharacteristic. In one aspect, a plant has “all but one, two, or threeof the physiological and morphological characteristics” of areferred-to-plant, or “all the physiological and morphologicalcharacteristics” of Table 1 “all or all but one, two, or three of thephysiological and morphological characteristics” of Table 1.

The physiological and/or morphological characteristics mentioned aboveare commonly evaluated at significance levels of 1%, 5% or 10% if theyare numerical, or for having an identical degree (or type) if notnumerical, if measured under the same environmental conditions. Forexample, a progeny plant or a Single Locus Converted Plant or a mutatedplant of pepper variety NUN 70064 PPH, may have one or more (or all) ofthe physiological and/or morphological characteristics of said varietylisted in Table 1, as determined at the 5% significance level (i.e.,p<0.05) when grown under the same environmental conditions.

“Distinguishing characteristics” or “distinguishing morphological and/orphysiological characteristics” refers herein to the characteristicswhich distinguish (i.e., are different) between the new pepper varietyfrom other pepper varieties, such as the Reference Variety when grownunder the same environmental conditions. When comparing pepper varietyNUN 70064 PPH with different varieties, the distinguishingcharacteristics may be different. In one aspect, the distinguishingcharacteristics may therefore include at least one, two, three or more(or all) of the characteristics listed in Table 1. All numericaldistinguishing characteristics are statistically significantly differentat p<0.05 between pepper variety NUN 70064 PPH and the other variety,e.g., the Reference Variety.

Thus, a pepper plant “comprising the distinguishing characteristics ofpepper variety NUN 70064 PPH” (such as a progeny plant) refers herein toa plant which does not differ significantly from said variety in thedistinguishing characteristics above, Therefore, in one aspect, a plant(such as a progeny plant of variety NUN 70064 PPH) is provided whichdoes not differ significantly from pepper variety NUN 70064 PPH in thedistinguishing characteristics above.

Similarity and differences between two different plant lines orvarieties can be determined by comparing the number of morphologicaland/or physiological characteristics (e.g. the characteristics as listedin Table 1) that are the same (i.e., statistically not significantlydifferent) or that are different (i.e., statistically significantlydifferent) between the two plant lines or varieties when grown under thesame environmental conditions. A numerical characteristic is consideredto be “the same” when the value for a numeric characteristic is notsignificantly different at the 1% (p<0.01) or 5% (p<0.05) significancelevel, using T-test, a standard method known to the skilled person.Non-numerical or “degree” or “type” characteristic is considered “thesame” when the values have the same “degree” or “type” when scored usingUSDA and/or UPOV descriptors, if the plants are grown under the sameenvironmental conditions.

In one aspect, a statistical analysis of the quantitativecharacteristics showing the degree of significance may be provided.Statistical significance is the likelihood that a relationship betweentwo or more variables is caused by something other than chance, i.e.,that the differences in the means for quantitative characteristics ofthe plant of pepper variety NUN 70064 PPH and the Reference Variety aresignificant due to chance. For the purpose of proving differences ordistinction between pepper variety NUN 70064 PPH and the ReferenceVariety, a p-value of 5% or 0.05 or lower is considered statisticallysignificant. This means that there is only a 5% probability that theobserved result could have happened just by chance or random variation.

The statistical analysis is drawn from a small sample of at least 15plants or plant parts of pepper variety NUN 70064 PPH and the ReferenceVariety. Statistical points or parameters such as mean, minimum, median,maximum, and standard deviation are collected from the sample data toanalyze where the average is, how varied the data set is, and whetherthe data is skewed. For the purpose of determining whether the result ofthe data set is statistically significant, a T-Test is used, astatistical tool for proving significance in the means of the two groups(e.g., pepper variety NUN 70064 PPH and the Reference Variety) at 5%significance level (a p-value of 5% or 0.05).

The term “traditional breeding techniques” encompasses herein crossing,selfing, selection, doubled haploid production, embryo rescue,protoplast fusion, marker assisted selection, mutation breeding, etc. asknown to the breeder (e.g., methods other than geneticmodification/transformation/transgenic methods), by which, for example,a genetically heritable trait can be transferred from one pepper line orvariety to another.

“Variety” or “cultivar” means a plant grouping within a single botanicaltaxon of the lowest known rank.

A “plant line” is for example a breeding line which can be used todevelop one or more varieties. A breeding line is typically highlyhomozygous.

“Hybrid variety” or “F1 hybrid” refers to the seeds harvested fromcrossing two inbred (nearly homozygous) parental lines. For example, thefemale parent is pollinated with pollen of the male parent to producehybrid (F1) seeds on the female parent.

“Tissue culture” or “cell culture” refers to a composition comprisingisolated cells of the same or a different type or a collection of suchcells organized into parts of a plant. Tissue culture of various tissuesof pepper and regeneration of plants therefrom is well known and widelypublished (see, e.g., Sang-Gu et al. (1988), Plant Cell, Tissue andOrgan Culture 12: 67-74; Kothari et al., (2010) Biotechnology Advances28: 35-48). Similarly, the skilled person is well-aware how to prepare a“tissue culture” or “cell culture.”

“Regeneration” refers to the development of a plant from cell culture ortissue culture or vegetative propagation.

“Vegetative propagation”, “vegetative reproduction” or “clonalpropagation” are used interchangeably herein and mean a method of takinga part of a plant and allowing that plant part to form at least roots,and also refer to the plant or plantlet obtained by that method.Optionally, the vegetative propagation is grown into a mature plant. Theskilled person is aware of what plant parts are suitable for use in themethod.

“Selfing” refers to self-pollination of a plant, i.e., the transfer ofpollen from the anther to the stigma of the same plant.

“Crossing” refers to the mating of two parent plants. The termencompasses “cross-pollination” and “selfing”.

“Cross-pollination” refers to the fertilization by the union of twogametes from different plants.

“Backcrossing” is a traditional breeding technique used to introduce atrait into a plant line or variety. The plant containing the trait iscalled the donor plant and the plant into which the trait is transferredis called the recurrent parent. An initial cross is made between thedonor parent and the recurrent parent to produce a progeny plant.Progeny plants which have the trait are then crossed to the recurrentparent. After several generations of backcrossing and/or selfing therecurrent parent comprises the trait of the donor. The plant generatedin this way may be referred to as a “single trait converted plant”. Thetechnique can also be used on a parental line of a hybrid.

“Progeny” as used herein refers to a plant obtained from a plantdesignated NUN 70064 PPH. A progeny may be obtained by regeneration ofcell culture or tissue culture or parts of a plant of said variety orselfing of a plant of said variety or by producing seeds of a plant ofsaid variety. In further aspects, progeny may also encompass plantsobtained from crossing of at least one plant of said variety withanother pepper plant of the same variety or another variety or(breeding) line, or with wild pepper plants. A progeny may comprise amutation or a transgene. A first generation progeny” or is the progenydirectly derived from, obtained from, obtainable from or derivable fromthe parent plant by, e.g., traditional breeding methods (selfing and/orcross-pollinating) or regeneration (optionally combined withtransformation and mutation). Thus, a plant of variety NUN 70064 PPH, isthe male parent, the female parent or both of a first generation progenyof that variety or both of a first generation progeny of pepper varietyNUN 70064 PPH. Progeny may have all the physiological and morphologicalcharacteristics of variety NUN 70064 PPH when grown under the sameenvironmental conditions and/or progeny may have (be selected forhaving) one or more of the distinguishing characteristics of pepper ofthe disclosure. Using common breeding methods such as backcrossing orrecurrent selection, mutation or transformation, one or more specificcharacteristics may be introduced into said variety, to provide or aplant comprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of variety NUN 70064 PPH.

The terms “gene converted” or “conversion plant” or “single locusconverted plant” in this context refer to pepper plants which aredeveloped by traditional breeding techniques e.g., backcrossing or viagenetic engineering or through mutation breeding, wherein essentiallyall of the desired morphological and physiological characteristics ofthe parent variety or line are recovered, in addition to the one or moregenes transferred into the parent via e.g., backcrossing technique(optionally including reverse breeding or reverse synthesis of breedinglines). It is understood that only the addition of a furthercharacteristic (e.g., addition of gene conferring a furthercharacteristic, such as a disease resistance gene), but also thereplacement/modification of an existing characteristic by a differentcharacteristic is encompassed herein.

Likewise, a “Single Locus Converted (Conversion) Plant” refers to aplant developed by plant breeding techniques comprising or consisting ofmutation and/or by genetic transformation and/or by traditional breedingtechniques, such as backcrossing, wherein essentially all of the desiredmorphological and physiological characteristics of a pepper variety arerecovered in addition to the characteristics of the single locus havingbeen transferred into the variety via the backcrossing technique. Incase of a hybrid, the gene may be introduced in the male or femaleparental line.

“Transgene” or “chimeric” gene” refers to a genetic locus comprising aDNA sequence which has been introduced into the genome of the plant bytransformation. A plant comprising a transgene stably integrated intoits genome is referred to as “transgenic plant.”

As used herein, the terms “resistance” and “tolerance” are usedinterchangeably to describe plants that show no symptoms orsignificantly reduced symptoms to a specified biotic pest, pathogen,abiotic influence or environmental condition compared to a susceptibleplant. These terms are optionally also used to describe plants showingsome symptoms but that are still able to produce marketable product withan acceptable yield.

“Average” refers herein to the arithmetic mean.

The term “mean” refers to the arithmetic mean of several measurements.The skilled person understands that the appearance of a plant depends tosome extent on the growing conditions of said plant. The mean, if notindicated otherwise within this application, refers to the arithmeticmean of measurements on at least 15 different, randomly selected plantsof a variety or line.

DETAILED DESCRIPTION OF VARIOUS ASPECTS OF THE DISCLOSURE

The disclosure relates to a plant of variety NUN 70064 PPH, wherein arepresentative sample of seeds of said variety has been deposited underthe Budapest Treaty, with Accession Number NCIMB ______. NUN 70064 PPHis a jalapeno pepper variety for the fresh market and is suitable forgrowing in the open field.

The disclosure also provides a pepper plant, or part thereof, having allof the physiological and morphological characteristics of pepper varietyNUN 70064 PPH when grown under the same environmental conditions.

In another aspect, pepper variety NUN 70064 PPH or a progeny thereofcomprises resistance to Tomato Spotted Wilt Virus (TSWV), measuredaccording to UPOV standards described in TG/76/8.

The disclosure also provides a plant of variety NUN 70064 PPH, or a partthereof, or a progeny plant thereof, comprising all of the followingmorphological and/or physiological characteristics (i.e., averagevalues, as indicated on the USDA Objective description of variety—pepper(unless indicated otherwise)) as shown in Table 1, determined at 5%significance level for numerical characteristics and determined by typeor degree for plants for non-numerical characteristics when grown underthe same environmental conditions. A part of this plant is alsoprovided.

The disclosure further provides a pepper plant which does not differfrom the physiological and morphological characteristics of the plant ofvariety NUN 70064 PPH as determined at the 1%, 2%, 3%, 4% or 5%significance level when grown under the same environmental conditions.In a particular aspect, the plants are measured in the same trial (e.g.,the trial is conducted as recommended by the USDA or UPOV). Thedisclosure also comprises a part of said plant, preferably a fruit orpart thereof.

The morphological and/or physiological differences between two differentindividual plants of the disclosure (e.g., between pepper variety NUN70064 PPH, and a progeny thereof) or between a plant of variety NUN70064 PPH or progeny of said variety, or a plant having all, or all but1, 2, or 3, of the physiological and morphological characteristics ofvariety NUN 70064 PPH (or all, or all but 1, 2, or 3 of thecharacteristics as listed in Table 1) and another known variety caneasily be established by growing said variety next to each other or nextto the other variety (in the same field, under the same environmentalconditions), preferably in several locations which are suitable for saidpepper cultivation, and measuring morphological and/or physiologicalcharacteristics of a number of plants (e.g., to calculate an averagevalue and to determine the variation range/uniformity within thevariety). For example, trials can be carried out in Acampo CA, USA (N 38degrees 07′261″/W 121 degrees 18′ 807″, USA), whereby variouscharacteristics, for example, maturity, days from seeding to harvest,plant habit, plant attitude, leaf shape, leaf color, blistering, numbersof flowers per leaf axil, number of calyx lobes, number of petals, fruitgroup, immature fruit color, mature fruit color, pungency, flavor, fruitglossiness, fruit size, fruit shape, average number of fruits per plant,seed size, seed weight, anthocyanin level, disease resistance, insectresistance, can be measured and directly compared for species of pepper.

Thus, the disclosure comprises pepper plant having one, two, or threephysiological and/or morphological characteristics which are differentfrom those of the plant of variety NUN 70064 PPH and which otherwise hasall the physiological and morphological characteristics of said variety,when determined at the 5% significance level for numericalcharacteristics and determined by type or degree for non-numericalcharacteristics for plants grown under the same environmentalconditions. In a particular aspect, the different characteristic(s)is/are result of breeding with pepper variety NUN 70064 PPH andselection of progeny plant comprising 1, 2, or 3 characteristics whichare different than in pepper variety NUN 70064 PPH. In another aspect,the different characteristic is the result of a mutation (e.g.,spontaneous mutation or a human induced mutation through e.g., targetedmutagenesis or traditional mutagenesis such as chemically or radiationinduced mutagenesis), or it is the result of transformation.

The disclosure also relates to a seed of pepper variety NUN 70064 PPH,wherein a representative sample of said seed has been deposited underthe Budapest Treaty with Accession Number NCIMB ______.

In another aspect, a seed of hybrid variety NUN 70064 PPH is obtainableby crossing the male parent of pepper variety NUN 70064 PPH with thefemale parent pepper variety NUN 70064 PPH and harvesting the seedsproduced on the female parent. The resultant seeds of said variety canbe grown to produce plants of said variety.

In another aspect, the disclosure provides a plant grown from a seed ofpepper variety NUN 70064 PPH and plant part thereof.

The disclosure also provides a pepper fruit produced on a plant grownfrom a seed of pepper variety NUN 70064 PPH.

In another aspect, the disclosure provides for a pepper plant part ofvariety NUN 70064 PPH, preferably a fruit, a representative sample ofseed from said variety has been deposited under the Budapest Treaty withAccession Number NCIMB ______.

Also provided is a plant of pepper variety NUN 70064 PPH, or a fruit orother plant part thereof, produced from a seed, wherein a representativesample of said seeds has been deposited under the Budapest Treaty withAccession Number NCIMB ______.

Also a plant part obtained from variety NUN 70064 PPH, is provided,wherein said plant part is: a fruit, a harvested fruit, a part of afruit, a leaf, a part of a leaf, pollen, an ovule, a cell, a petiole, ashoot or a part thereof, a stem or a part thereof, a root or a partthereof, a root tip, a cutting, a seed, a part of a seed, seed coat oranother maternal tissue which is part of a seed grown on said varieties,a hypocotyl, a cotyledon, a scion, a stock, a rootstock, a pistil, ananther, and a flower or a part thereof. Fruits are particularlyimportant plant parts. Fruits may be parthenocarpic, or seedless, orcontain immature and/or nonviable seeds.

In a further aspect, the plant part obtained from variety NUN 70064 PPHis a cell, optionally a cell in a cell or tissue culture. That cell maybe grown into a plant of variety NUN 70064 PPH. A part of a variety ofNUN 70064 PPH (or of progeny variety NUN 70064 PPH, or of a plant havingall physiological and/or morphological characteristics but one, two, orthree which are different from those of variety NUN 70064 PPH) furtherencompasses any cells, tissues, organs obtainable from the seedlings orplants in any stage of maturity.

The disclosure also provides a tissue or cell culture comprising cellsof pepper variety NUN 70064 PPH. Such a tissue culture can, for example,be grown on plates or in liquid culture, or be frozen for long termstorage. The cells of pepper variety NUN 70064 PPH, used to start theculture can be selected from any plant part suitable for vegetativereproduction, or in a particular aspect can be one or more of an embryo,meristem, a cotyledon, a hypocotyl, pollen, a leaf, an anther, a root, aroot tip, a pistil, a petiole, a flower, a fruit, seed, and/or a stem.In another particular aspect, the tissue culture does not containsomaclonal variation or has reduced somaclonal variation. The skilledperson is familiar with methods to reduce or prevent somaclonalvariation, including regular reinitiation.

In one aspect, the disclosure provides a pepper plant regenerated fromthe tissue or cell culture of pepper variety NUN 70064 PPH, wherein theregenerated plant is not significantly different from pepper variety NUN70064 PPH in all, or all but one, two, or three, of the physiologicaland morphological characteristics, e.g. determined at the 5%significance level for numerical characteristics and determined by typeor degree for non-numerical characteristics when grown under the sameenvironmental conditions. Optionally, the plant has one, two, or threethe physiological and morphological characteristics that are affected bya mutation or by transformation. In another aspect, the disclosureprovides a pepper plant regenerated from the tissue or cell culture ofpepper variety NUN 70064 PPH, wherein the plant has all of thephysiological and morphological characteristics of said variety, e.g.,determined at the 5% significance level for numerical characteristicsand determined by type or degree for non-numerical characteristics whengrown under the same environmental conditions. Similarity or differenceof a characteristic is determined by measuring the characteristics of arepresentative number of plants grown under the same environmentalconditions, determining whether type/degree characteristics are the sameand determining whether numerical characteristics are different at the5% significance level.

Pepper variety NUN 70064 PPH, or its progeny, or a plant having allphysiological and/or morphological characteristics but one, two, orthree which are different from those of pepper variety NUN 70064 PPH canalso be reproduced using vegetative reproduction methods. Therefore, thedisclosure provides for a method of producing a plant, or a partthereof, of variety NUN 70064 PPH, comprising vegetative propagation ofsaid variety. Vegetative propagation comprises regenerating a wholeplant from a plant part of variety NUN 70064 PPH, (or from a progeny ofsaid variety or from or a plant having all physiological and/ormorphological characteristics of said variety but one, two, or threedifferent characteristics), such as a cutting, a cell culture or atissue culture.

The disclosure also concerns methods of vegetatively propagating a partof the plant of pepper variety NUN 70064 PPH. In certain aspects, themethod comprises: (a) collecting tissue or cells capable of beingpropagated from a plant described herein; (b) cultivating said tissue orcells to obtain proliferated shoots; and (c) rooting said proliferatedshoots, to obtain rooted plantlets. Steps (b) and (c) may also bereversed, i.e. first cultivating said tissue to obtain roots and thencultivating the tissue to obtain shoots, thereby obtaining rootedplantlets. The rooted plantlets may then be further grown, to obtainplants. In one aspect, the method further comprises step (d) growingplants from said rooted plantlets. Therefore, the method also comprisesregenerating a whole plant from said part of NUN 70064 PPH. In aparticular aspect, the part of the plant to be propagated is a cutting,a cell culture or a tissue culture.

The disclosure also provides for a vegetatively propagated plant ofvariety NUN 70064 PPH (or from progeny of said variety or from or aplant having all but one, two or three physiological and/ormorphological characteristics of said variety), wherein the plant hasall of the morphological and physiological characteristics of varietyNUN 70064 PPH, e.g., determined at the 5% significance level fornumerical characteristics and determined by type or degree for plantsgrown under the same environmental conditions. In another aspect, thepropagated plant has all but one, two, or three of the morphological andphysiological characteristics of variety NUN 70064 PPH, e.g., determinedat the 5% significance level for numerical characteristics anddetermined by type or degree for non-numerical characteristics forplants grown under the same environmental conditions. A part of saidpropagated plant or said propagated plant with one, two or threedifferences is also included. In another aspect, the propagated planthas all or all but, 1, 2, or 3 of the physiological and morphologicalcharacteristics of pepper variety NUN 70064 PPH (e.g., as listed inTable 1).

In another aspect, the disclosure provides a method for producing apepper plant part, preferably a fruit, comprising growing a plant ofvariety NUN 70064 PPH until it sets at least one fruit, and collectingthe fruit. Preferably, the fruit is collected at harvest maturity. Inanother aspect, the fruit is collected when the seed is ripe.

In another aspect, a plant of variety NUN 70064 PPH can be produced byseeding directly in the soil (e.g., field) or by germinating the seedsin controlled environment conditions (e.g., greenhouses) and optionallythen transplanting the seedlings into the field. For example, the seedcan be sown into prepared seed beds where they will remain for theentire production the crop (see, Hartz, et. al, University of CaliforniaDivision of Agriculture and Natural Resources, Publication 7217, 1-4).Stakes and plastic mulches may also be used for peppers for the freshthe market, particularly, when peppers are to be harvested at maturefruit color and to promote earliness and yield. On the other hand, nostake or mulch is used for processing peppers. Moreover, pepper can alsobe grown entirely in greenhouses or tunnels.

In another aspect, the disclosure provides a progeny plant of varietyNUN 70064 PPH, such as a progeny plant obtained by further breeding thatvariety. Further breeding with the variety includes selfing that varietyone or more times and/or cross-pollinating that variety with anotherpepper plant or variety one or more times. In particular, the disclosureprovides for a progeny plant that retains all the morphological andphysiological characteristics of pepper variety NUN 70064 PPH, or, inanother aspect, a progeny plant that retains all, or all but one, two,or three, of the morphological and physiological characteristics ofvariety NUN 70064 PPH, optionally all or all but one, two, or three ofthe characteristics as listed in Table 1, e.g., determined at the 5%significance level for numerical characteristics and determined by typeor degree for non-numerical characteristics when grown under the sameenvironmental conditions. In a particular aspect, the progeny is a firstgeneration progeny, i.e., the ovule or the pollen (or both) used in thecrossing is an ovule or pollen of variety NUN 70064 PPH, where thepollen comes from an anther of and the ovule comes from an ovary ofvariety NUN 70064 PPH and the ovule comes from an ovary of peppervariety NUN 70064 PPH.

In still another aspect, the disclosure provides a method of producing apepper plant, comprising crossing a plant of pepper variety NUN 70064PPH with a second pepper plant at least once, allowing seed to developand optionally harvesting said progeny seed. The skilled person canselect progeny from said crossing. Optionally, the progeny (grown fromthe progeny seed) is crossed twice, thrice, or four, five, six or seventimes, and allowed to set seed. In one aspect of the disclosure, thefirst “crossing” further comprises planting seeds of a first and asecond parent pepper plant, often in proximity so that pollination willoccur; for example, mediated by insect vectors. Alternatively, pollencan be transferred manually. Where the plant is self-pollinated,pollination may occur without the need for direct human interventionother than plant cultivation. After pollination the plant can produceseed.

The disclosure also provides a method for collecting pollen, comprisingcollecting pollen from a pepper variety NUN 70064 PPH. Alternatively,the method comprises growing a plant of variety NUN 70064 PPH until atleast one flower contains pollen and collecting the pollen. Inparticular aspect, the pollen is collected when it is mature or ripe. Asuitable method for collecting pollen comprises collecting anthers orthe part of the anther that contains pollen, for example by cutting theanther or the part of the anther off. Pollen can be collected in acontainer. Optionally, collected pollen can be used to pollinate apepper flower.

In yet another aspect, the disclosure provides a method of producing aplant, comprising selfing a plant of variety NUN 70064 PPH one or moretimes, and selecting a progeny plant from said selfing. In one aspect,the progeny plant retains all or all but one, two, or three of themorphological and physiological characteristics of variety NUN 70064 PPHwhen grown under the same environmental conditions. In a differentaspect, the progeny plant comprises all (or all but one, two, or three)of the physiological and morphological characteristic of variety NUN70064 PPH of Table 1.

The disclosure also provides a method for developing a pepper plant in apepper breeding program, using a pepper plant described herein, or itsparts as a source of plant breeding material. Suitable plant breedingtechniques are recurrent selection, backcrossing, pedigree breeding,mass selection, mutation breeding and/or genetic marker enhancedselection. For example, in one aspect, the method comprises crossingpepper variety NUN 70064 PPH, or progeny of said variety, or a plantcomprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of variety NUN 70064 PPH (e.g., as listedin Table 1), with a different pepper plant, and wherein one or moreoffspring of the crossing are subject to one or more plant breedingtechniques: recurrent selection, backcrossing, pedigree breeding, massselection, mutation breeding and genetic marker enhanced selection (seee.g., Martin et al. 2008, Australian Journal of Crop Science 1(2):43-46). For breeding methods in general (see, e.g., Principles of PlantGenetics and Breeding, 2007, George Acquaah, Blackwell Publishing,ISBN-13: 978-1-4051-3646-4).

In one aspect, pedigree selection is used as a breeding method fordeveloping a pepper variety. Pedigree selection is also known as the“Vilmorin System of Selection,” see, e.g., Allard, John Wiley & Sons,Inc., 1999, pp. 64-67. In general, selection is first practiced among F2plants. In the next season, the most desirable F3 lines are firstidentified, then desirable F3 plants within each line are selected. Thefollowing season and in all subsequent generations of inbreeding, themost desirable families are identified first, then desirable lineswithin the selected families are chosen. A family refers to lines thatwere derived from plants selected from the same progeny from thepreceding generation.

Thus, progeny in connection with pedigree selection are either thegeneration (seeds) produced from the first cross (F1) or selfing (S1),or any further generation produced by crossing and/or selfing (F2, F3,etc.) and/or backcrossing (BC1, BC2, etc.) one or more selected plantsof the F1 and/or S1 and/or BC1 generation (or plants of any furthergeneration, e.g., F2) with another pepper plant (and/or with a wildrelative of pepper). Progeny may have all the physiological andmorphological characteristics of pepper variety NUN 70064 PPH when grownunder the same environmental conditions and/or progeny may have (beselected for having) one or more of the distinguishing characteristicsof pepper variety NUN 70064 PPH.

In yet a further aspect, the disclosure provides for a method ofproducing a pepper plant. The method comprises crossing a plant ofvariety NUN 70064 PPH, or a plant comprising all but 1, 2, or 3 of themorphological and physiological characteristics of pepper variety NUN70064 PPH (e.g., as listed in Table 1), or a progeny plant thereof,either as male or as female parent, with a second pepper plant (or awild relative of pepper) one or more times, and/or selfing a pepperplant of variety NUN 70064 PPH, or a progeny plant thereof, one or moretimes, and selecting progeny from said crossing and/or selfing. Thesecond pepper plant may, for example, be a line or variety of thespecies species Capsicum annuum, C. frutecens, C. baccatum, C. chinense,or other Capsicum species.

In a further aspect, pepper variety NUN 70064 PPH is used in crosseswith other, different, pepper varieties to produce first generation (F1)pepper hybrid seeds and plants with superior characteristics. In aparticular aspect, the disclosure provides a pepper seed and a plantproduced by crossing a first parent pepper plant with a second parentpepper plant, wherein at least one of the first or second parent pepperplant is pepper variety NUN 70064 PPH. In another aspect, the pepperseed and plant produced are the first filial generation (F1) pepper seedand plants produced by crossing the plant of pepper variety NUN 70064PPH with another pepper plant.

The morphological and physiological characteristics (and thedistinguishing characteristics) of pepper variety NUN 70064 PPH areprovided, e.g., Table 1. Encompassed herein is also a plant obtainablefrom pepper variety NUN 70064 PPH (e.g., by selfing and/or crossingand/or backcrossing with said variety and/or progeny of said variety)comprising all or all but one, two, or three of the physiological andmorphological characteristics of variety NUN 70064 PPH e.g., as listedin Table 1 when determined at the 5% significance level for numericalcharacteristics or determined by type or degree for non-numericalcharacteristics when grown under the same environmental conditionsand/or comprising one or more (or all; or all except one, two, or three)characteristics when grown under the same environmental conditions. Themorphological and/or physiological characteristics may vary somewhatwith variation in the environment (such as temperature, light intensity,day length, humidity, soil, fertilizer use), which is why a comparisonunder the same environmental conditions is preferred. Colors can best bemeasured using the Royal Horticultural Society Chart.

In another aspect, the disclosure provides a method of producing a plantderived from pepper variety NUN 70064 PPH, comprising crossing a plantof pepper variety NUN 70064 PPH either as a male or female parent with asecond plant or selfing pepper variety NUN 70064 PPH or vegetativereproduction of pepper variety NUN 70064 PPH and collecting seeds fromsaid crossing or selfing or regenerating a whole plant from thevegetable cell-or tissue culture. Also provided are seeds and/or plantsobtained by this method. All plants produced using pepper variety NUN70064 PPH as a parent are within the scope of the disclosure, includingplant parts derived from pepper variety NUN 70064 PPH.

In further aspects, the method comprises growing a progeny plant of asubsequent generation and crossing the progeny plant of a subsequentgeneration with itself or a second plant and repeating the steps foradditional 3-10 generations to produce a plant derived from peppervariety NUN 70064 PPH. The plant derived from pepper variety NUN 70064PPH may be an inbred line and the aforementioned repeating crossingsteps may be defined as comprising sufficient inbreeding to produce theinbred line. By selecting plants having one or more desirable traits ofthe line as well as potentially other selected traits.

The disclosure provides for methods of producing plants which retain allthe morphological and physiological characteristics of a plant ofvariety NUN 70064 PPH. The disclosure provides also for methods ofproducing a plant comprising all but 1, 2, or 3 or more of themorphological and physiological characteristics of variety NUN 70064 PPH(e.g., as listed in Table 1), but which are still genetically closelyrelated to said variety. The relatedness can, for example, be determinedby fingerprinting techniques (e.g., making use of isozyme markers and/ormolecular markers such as Single-nucleotide polymorphism (SNP) markers,amplified fragment length polymorphism (AFLP) markers, microsatellites,minisatellites, Random Amplified Polymorphic DNA (RAPD) markers,restriction fragment length polymorphism (RFLP) markers and others). Aplant is “closely related” to variety NUN 70064 PPH, if its DNAfingerprint is at least 80%, 90%, 95% or 98% identical to thefingerprint of said variety. In a particular aspect, AFLP markers areused for DNA fingerprinting (see, e.g., Vos et al. 1995, Nucleic AcidResearch 23: 4407-4414). A closely related plant may have a Jaccard'sSimilarity index of at least about 0.8, preferably at least about 0.9,0.95, 0.98 or more (see, e.g., Ince, et. al., 2010, Biochem Genet48:83-95). The disclosure also provides a plant and a variety obtainedor selected by applying these methods on pepper variety NUN 70064 PPH.Such a plant may be produced by crossing and/or selfing, oralternatively, a plant may simply be identified and selected amongstplants of said variety, or progeny of said variety, e.g., by identifyinga variant within pepper variety NUN 70064 PPH, or within progeny of saidvariety (e.g., produced by selfing) which variant differs from saidvariety in one, two, or three of the morphological and/or physiologicalcharacteristics (e.g., in one, two, or three distinguishingcharacteristics), e.g., as listed in Table 1. In one aspect, thedisclosure provides a pepper plant having a Jaccard's Similarity indexwith pepper variety NUN 70064 PPH of at least 0.8, e.g., at least 0.85,0.9, 0.95, 0.98 or even at least 0.99.

In another aspect, the disclosure provides a pepper plant comprisinggenomic DNA having at least 95%, 96%, 97%, 98% or 99% sequence identitycompared to the genomic DNA sequence of a plant of variety NUN 70064 PPHas deposited under Accession Number NCIMB ______. In some aspects, thepepper plant further comprises all or all but 1, 2, or 3 of themorphological and physiological characteristics of variety NUN 70064 PPH(e.g., as listed in Table 1). In other aspects, the pepper plant is ahybrid derived from a seed or plant of variety NUN 70064 PPH.

For the purpose of this disclosure, the “sequence identity” ofnucleotide sequences, expressed as a percentage, refers to the number ofpositions in the two optimally aligned sequences which have identicalresidues (×100) divided by the number of positions compared. A gap,i.e., a position in the pairwise alignment where a residue is present inone sequence but not in the other, is regarded as a position withnon-identical residues. A pairwise global sequence alignment of twonucleotide sequences is found by aligning the two sequences over theentire length according to the Needleman and Wunsch global alignmentalgorithm described in Needleman and Wunsch, 1970, J. Mol. Biol.48(3):443-53). A full implementation of the Needleman-Wunsch globalalignment algorithm is found in the needle program in The EuropeanMolecular Biology Open Software Suite (see, e.g., EMBOSS, Rice et al.,Trends in Genetics June 2000, vol. 16, No. 6. pp. 276-277).

In another aspect, a plant of variety NUN 70064 PPH may also be mutated(by e.g., irradiation, chemical mutagenesis, heat treatment, etc.) andmutated seeds or plants may be selected in order to change one or morecharacteristics of said variety. Methods such as TILLING (TargetingInduced Local Lesions in Genomes) may be applied to pepper populationsin order to identify mutants.

Similarly, pepper variety NUN 70064 PPH may be transformed andregenerated, whereby one or more chimeric genes are introduced into thevariety or into a plant comprising all but 1, 2, 3, or more of themorphological and physiological characteristics (e.g., as listed inTable 1). Many useful traits can be introduced into pepper variety NUN70064 PPH by e.g., crossing pepper variety NUN 70064 PPH with atransgenic pepper plant comprising a desired transgene as well as bydirectly introducing a transgene into pepper variety NUN 70064 PPH bygenetic transformation techniques.

Transformation can be carried out using standard methods, such asAgrobacterium tumefaciens mediated transformation or biolistics,followed by selection of the transformed cells and regeneration intoplants. A desired trait (e.g., gene(s) conferring pest or diseaseresistance, herbicide, fungicide or insecticide tolerance, etc.) can beintroduced into pepper variety NUN 70064 PPH, or progeny of saidvariety, by transforming said variety or progeny of said variety with atransgene that confers the desired trait, wherein the transformed plantretains all or all but one, two or three of the phenotypic and/ormorphological and/or physiological characteristics of variety NUN 70064PPH, or the progeny of said variety and contains the desired trait.

Any pest or disease resistance genes may be introduced into a plant ofvariety NUN 70064 PPH, progeny of said variety or into a plantcomprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of variety NUN 70064 PPH (e.g., as listedin Table 1). Resistance to one or more of the following diseases orpests may be introduced into plants of the disclosure: Cucumber MosaicVirus, Curly Top Virus, Pepper Mild Mottle Virus, Potato Y Virus,Tobacco Etch Virus, Tobacco Mosaic Virus, Tomato Spotted Wilt Virus,Anthracnose (Gloeosporium piperatum), Bacterial Spot (Xanthomonasvesicatoria), Cercospora Leaf Spot (Cercospora capsici), Nematode(Meloidogyne incognita acrita), Phytophthora Root Rot (Phytophthoracapsici), Ripe Rot (Vermicularia capsici), Southern Blight (Sclerotiumrolfsii) and/or Verticillium Wilt (Verticillium dahliae). Otherresistance genes, against pathogenic viruses, fungi, bacteria,nematodes, insects or other pests may also be introduced.

Genetic transformation may, therefore, be used to insert a selectedtransgene into the pepper plants of the disclosure described herein ormay, alternatively, be used for the preparation of transgenic pepperplants which can be used as a source of the transgene(s), which can beintroduced into pepper variety NUN 70064 PPH by e.g., backcrossing. Agenetic trait which has been engineered into the genome of a particularpepper plant may then be moved into the genome of another pepper plant(e.g., another variety) using traditional breeding techniques which arewell known in the art. For example, backcrossing is commonly used tomove a transgene from a transformed pepper variety into an alreadydeveloped pepper variety and the resulting backcross conversion plantwill then comprise the transgene(s).

Any DNA sequences, whether from a different species or from the samespecies, which are inserted into the genome using transformation, arereferred to herein collectively as “transgenes.” A “transgene” alsoencompasses antisense, or sense and antisense sequences capable of genesilencing. Thus, the disclosure also relates to transgenic plants ofpepper variety NUN 70064 PPH. In some aspects, a transgenic plant ofpepper variety NUN 70064 PPH may contain at least one transgene butcould also contain at least 1, 2, 3, 4, or more transgenes.

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of, or operatively linked to aregulatory element active in plant cells (e.g., promoter). Theexpression vector may contain one or more such operably linkedgene/regulatory element combinations. The vector may be in the form of aplasmid and can be used alone or in combination with other plasmids toprovide transformed pepper plants using transformation methods toincorporate transgenes into the genetic material of the pepper plant(s).Transformation can be carried out using standard methods, such asAgrobacterium tumefaciens mediated transformation, electroporation,biolistics particle delivery system, or microprojectile bombardment,followed by selection of the transformed cells and regeneration intoplants.

Plants can also be genetically engineered, modified, or manipulated toexpress various phenotypes of horticultural interest. Through thetransformation of pepper, the expression of genes can be altered toenhance disease resistance, insect resistance, herbicide resistance,stress tolerance, horticultural quality, and other traits.Transformation can also be used to insert DNA sequences which control orhelp control male sterility or fertility restoration. DNA sequencesnative to pepper as well as non-native DNA sequences can be transformedinto pepper and used to alter levels of native or non-native proteins.Various promoters, targeting sequences, enhancing sequences, and otherDNA sequences can be inserted into the genome for the purpose ofaltering the expression of proteins. Reduction of the specific activityof specific genes (also known as gene silencing or gene suppression) isdesirable for several aspects of genetic engineering in plants.

Genome editing is another method recently developed to geneticallyengineer plants. Specific modification of chromosomal loci or targetedmutation can be done through sequence-specific nucleases (SSNs) byintroducing a targeted DNA double strand break in the locus to bealtered. Examples of SSNs that have been applied to plants are: fingernucleases (ZFNs), transcription activator-like effector nucleases(TALENs), engineered homing endonucleases or meganucleases, andclustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 (Cas9), see, e.g., Songstad, et.al., Critical Reviews in Plant Sciences, 2017, 36:1, 1-23.

Thus, the disclosure also provides a method of producing a pepper plantcomprising a desired trait, said method comprising mutating a plant, orplant part, of pepper variety NUN 70064 PPH and selecting a pepper plantcomprising the desired trait, wherein the mutated plant contains thedesired trait and otherwise retains all of the physiological andmorphological characteristics of pepper variety NUN 70064 PPH,optionally as described in Table 1, wherein a representative sample ofseed of said variety is deposited under Accession Number NCIMB ______.In a further aspect, the desired trait is yield, compact pepper, fruitquality, male sterility, herbicide tolerance, insect resistance, pestresistance, disease resistance, Powdery mildew resistance withoutnecrosis, environmental stress tolerance, modified carbohydratemetabolism, modified protein metabolism, or ripening.

In one aspect, the disclosure provides a method for inducing mutation inpepper variety NUN 70064 PPH, comprising:

-   -   a. exposing the seed, plant or plant part or cell of variety NUN        70064 PPH to a mutagenic compound or to radiation, wherein a        representative sample of seed of said pepper variety is        deposited under Accession Number NCIMB ______;    -   b. selecting the seed, plant or plant part or cell of variety        NUN 70064 PPH, having a mutation; and    -   c. optionally growing and/or multiplying the seed, plant or        plant part or cell of variety NUN 70064 PPH, having the        mutation.

The disclosure also provides a method of producing a pepper plant havinga desired trait, wherein the method comprises transforming the pepperplant with a transgene that confers the desired trait, wherein thetransformed plant contains the desired trait and otherwise retains allof the physiological and morphological characteristics of the plant ofvariety NUN 70064 PPH. Thus, a transgenic pepper plant is provided whichis produced by the method described above, wherein the plant otherwisehas all of the physiological and morphological characteristics of theplant of variety NUN 70064 PPH and the desired trait.

In another aspect, the disclosure provides a method of producing aprogeny of plant of variety NUN 70064 PPH, further comprising a desiredtrait, said method comprising transforming the plant of pepper varietyNUN 70064 PPH with at least one transgene that confers the desired traitand/or crossing the plant of pepper variety NUN 70064 PPH with atransgenic pepper plant comprising a desired transgene so that thegenetic material of the progeny that resulted from the cross containsthe desired transgene(s). Also encompassed is the progeny produced bythis method.

A desired trait (e.g., gene(s) conferring pest or disease resistance, ortolerance for protection, etc.) can be introduced into pepper varietyNUN 70064 PPH, or progeny of said variety, by transforming said varietyor progeny of said variety with a transgene that confers the desiredtrait, wherein the transformed plant retains all or all but one, two, orthree of the morphological and/or physiological characteristics ofpepper variety NUN 70064 PPH, or the progeny of said variety, andcontains the desired trait. In another aspect, the transformation ormutation confers a trait wherein the trait is yield, compact pepper,fruit quality, male sterility, herbicide tolerance, insect resistance,pest resistance, disease resistance, Powdery mildew resistance withoutnecrosis, environmental stress tolerance, modified carbohydratemetabolism, modified protein metabolism, or ripening. In a particularaspect, the specific transgene may be any known in the art or listedherein, including, a polynucleotide sequence conferring resistance toimidazolinone, sulfonylurea, glyphosate, glufosinate, triazine,benzonitrile, cyclohexanedione, phenoxy proprionic acid andL-phosphinothricin or a polynucleotide conferring resistance to CucumberMosaic Virus, Curly Top Virus, Pepper Mild Mottle Virus, Potato Y Virus,Tobacco Etch Virus, Tobacco Mosaic Virus, Tomato Spotted Wilt Virus,Anthracnose (Gloeosporium piperatum), Bacterial Spot (Xanthomonasvesicatoria), Cercospora Leaf Spot (Cercospora capsici), Nematode(Meloidogyne incognita acrita), Phytophthora Root Rot (Phytophthoracapsici), Ripe Rot (Vermicularia capsici), Southern Blight (Sclerotiumrolfsii) and/or Verticillium Wilt (Verticillium dahliae). Otherresistance genes, against pathogenic viruses, fungi, bacteria,nematodes, insects or other pests may also be introduced.

By crossing and/or selfing (one or more) single traits may be introducedinto the plant of variety NUN 70064 PPH (e.g., using backcrossingbreeding schemes), while retaining the remaining morphological andphysiological characteristics of said variety and/or while retaining oneor more or all distinguishing characteristics. A single trait convertedplant may thereby be produced. For example, disease resistance genes maybe introduced, genes responsible for one or more quality traits, yield,etc. Both single genes (e.g., dominant or recessive) and one or moreQTLs (quantitative trait loci) may be transferred into the plant ofpepper variety NUN 70064 PPH by breeding with said variety.

In another method, the step of introducing a single locus conversion inat least one of the parents comprises genetically transforming ormutating cells the parental line of pepper variety NUN 70064 PPH;growing the cells into a plant; and optionally selecting plants thatcontain the single locus conversion, the single trait conversion or thedesired trait.

In another aspect, the step of introducing a single locus conversion,single trait conversion, or a desired trait in at least one of theparent lines comprises:

-   -   a. crossing the parental line of pepper variety NUN 70064 PPH,        with a second pepper plant comprising the single locus        conversion, the single trait conversion or the desired trait;    -   b. selecting F1 progeny plants that contain the single locus        conversion, the single trait conversion or the desired trait;    -   c. crossing said selected progeny plants of step b) with the        parental line of step a), to produce a backcross progeny plant;    -   d. selecting backcross progeny plants comprising the single        locus conversion, the single trait conversion or the desired        trait and otherwise all or all but one, two, or three of the        morphological and physiological characteristics the parental        line of step a) to produce selected backcross progeny plants;        and    -   e. optionally repeating steps c) and d) one or more times in        succession to produce selected second, third or fourth or higher        backcross progeny plants comprising the single locus conversion,        the single trait conversion or the desired trait and otherwise        all or all but one, two, or three of the morphological and        physiological characteristics the parental line of step a) to        produce selected backcross progeny plants when grown in the same        environmental conditions.

The disclosure further relates to plants obtained by this method.

Alternatively, a single trait converted plant or single locus convertedplant may be produced by:

-   -   a. obtaining a cell or tissue culture of cells of pepper variety        NUN 70064 PPH;    -   b. genetically transforming or mutating said cells;    -   c. growing the cells into a plant; and    -   d. optionally selecting plants that contain the single locus        conversion, the single trait conversion, or the desired trait.

In another aspect, the disclosure provides a method of introducing asingle locus conversion or single trait conversion, or a desired traitinto pepper variety NUN 70064 PPH, comprising:

-   -   a. obtaining a combination of a parental lines of pepper variety        NUN 70064 PPH, optionally through reverse synthesis of breeding        lines;    -   b. introducing a single locus conversion in at least one of the        parents of step a; and    -   c. crossing the converted parent with the other parent of step a        to obtain seed of pepper variety NUN 70064 PPH.

In another aspect, the disclosure provides a method of introducing asingle locus conversion, or single trait conversion or a desired traitinto pepper variety NUN 70064 PPH, comprising introducing a single locusconversion, or single trait conversion, or a desired trait in at leastone of the parents of pepper variety NUN 70064 PPH; and crossing theconverted parent with the other parent of pepper variety NUN 70064 PPHto obtain seed of pepper variety NUN 70064 PPH.

In the above methods, wherein the single locus conversion concerns atrait, the trait may be yield or pest resistance or disease resistance.In one aspect, the trait is disease resistance and the resistance isconferred to Cucumber Mosaic Virus, Curly Top Virus, Pepper Mild MottleVirus, Potato Y Virus, Tobacco Etch Virus, Tobacco Mosaic Virus, TomatoSpotted Wilt Virus, Anthracnose (Gloeosporium piperatum), Bacterial Spot(Xanthomonas vesicatoria), Cercospora Leaf Spot (Cercospora capsici),Nematode (Meloidogyne incognita acrita), Phytophthora Root Rot(Phytophthora capsici), Ripe Rot (Vermicularia capsici), Southern Blight(Sclerotium rolfsii) and/or Verticillium Wilt (Verticillium dahliae).Other resistance genes, against pathogenic viruses, fungi, bacteria,nematodes, insects or other pests may also be introduced.

The disclosure also provides a plant having one, two, or threephysiological and/or morphological characteristics which are differentfrom those of pepper variety NUN 70064 PPH and which otherwise has allthe physiological and morphological characteristics of said variety,wherein a representative sample of seed of said pepper variety isdeposited under Accession Number NCIMB ______. In particular, variantswhich differ from pepper variety NUN 70064 PPH, in none, one, two, orthree of the characteristics mentioned in Table 1 are encompassed.

The disclosure also provides a pepper plant comprising at least a firstset of the chromosomes of pepper variety NUN 70064 PPH, a sample of seedof said variety is deposited under Accession Number NCIMB ______;optionally further comprising a single locus conversion or a mutation,wherein said plant has all of the morphological and physiologicalcharacteristics of the plant comprising at least a first set of thechromosomes of said variety. In another aspect, this single locusconversion confers a trait: yield, compact pepper, fruit quality,storage properties, color, male sterility, herbicide tolerance, insectresistance, pest resistance, disease resistance, environmental stresstolerance, modified carbohydrate metabolism, modified proteinmetabolism, or ripening.

In another aspect, the disclosure provides for a haploid plant and/or adoubled haploid plant of variety NUN 70064 PPH, or of a plant having allbut one, two, or three physiological and/or morphologicalcharacteristics of variety NUN 70064 PPH, or progeny of said variety.Haploid and doubled haploid (DH) plants can, for example, be produced bycell or tissue culture and chromosome doubling agents and regenerationinto a whole plant. DH production chromosome doubling may be inducedusing known methods, such as colchicine treatment or the like). In oneaspect, the method comprises inducing a cell or tissue culture with achromosome doubling agent and regenerating the cells or tissues into awhole plant.

In another aspect, the disclosure comprises a method for making doubledhaploid cells from haploid cells of variety NUN 70064 PPH, said methodcomprising doubling cells of with a doubling agent, such as a colchicinetreatment (see, e.g., Nikolova V, Niemirowicz-Szczytt K (1996) Acta SocBot Pol 65:311-317).

In yet another aspect, the disclosure provides for haploid plants and/ordoubled haploid plants derived from pepper variety NUN 70064 PPH that,when combined, make a set of parents of pepper variety NUN 70064 PPH.The haploid plant and/or the doubled haploid plant of NUN 70064 PPH canbe used in a method for generating parental lines of pepper variety NUN70064 PPH.

The disclosure also provides methods for determining the identity ofparental lines of plants described herein, in particular the identity ofthe female line. US2015/0126380, which is hereby incorporated byreference, relates to a non-destructive method for analyzing maternalDNA of a seed. In this method, the DNA is dislodged from the seed coatsurface and can be used to collect information on the genome of thematernal parent of the seed. This method for analyzing maternal DNA of aseed comprises contacting a seed with a fluid to dislodge DNA from theseed coat surface, and analyzing the DNA thus dislodged from the seedcoat surface using methods known in the art. The skilled person is thusable to determine whether a seed has grown on a plant of a plant ofvariety NUN 70064 PPH, is a progeny of said variety, because the seedcoat of the seed is a maternal tissue genetically identical to saidvariety. In one aspect, the disclosure relates to a seed coat comprisingmaternal tissue of pepper variety NUN 70064 PPH. In another aspect, thedisclosure relates to a pepper seed comprising a maternal tissue ofpepper variety NUN 70064 PPH. In another particular aspect, thedisclosure provides for a method of identifying the female parental lineof pepper variety NUN 70064 PPH by analyzing the seed coat of a seed ofthat variety. In another aspect, the skilled person can determinewhether a seed is grown on pepper variety NUN 70064 PPH by analyzing theseed coat of said seed.

Using methods known in the art such as “reverse synthesis of breedinglines” or “reverse breeding”, it is possible to produce parental linesfor a hybrid plant such as pepper variety NUN 70064 PPH. Thus, a skilledperson can take any individual heterozygous plant (called a“phenotypically superior plant” in Example 2 of US2015/0245570, herebyincorporated by reference in its entirety; pepper variety NUN 70064 PPHis such plant) and generate a combination of parental lines (reversebreeding parental lines) that, when crossed, produce pepper variety NUN70064 PPH. It is not necessary that the reverse breeding parental linesare identical to the original parental lines. Such breeding methods arebased on the segregation of individual alleles in the spores produced bya desired plant and/or in the progeny derived from the self-pollinationof that desired plant, and on the subsequent identification of suitableprogeny plants in one generation, or in a limited number of inbredcycles. Such a method is known from US2015/0245570 or from Wijnker etal., Nature Protocols Volume: 9, Pages: 761-772 (2014) DOI:doi:10.1038/nprot.2014.049. Thus, the disclosure provides a method forproducing parental lines for a hybrid organism (e.g., pepper variety NUN70064 PPH), comprising in one aspect: a) defining a set of geneticmarkers present in a heterozygous form (H) in a partially heterozygousstarting organism; b) producing doubled haploid lines from spores of thestarting organism; c) genetically characterizing the doubled haploidlines thus obtained for the said set of genetic markers to determinewhether they are present in a first homozygous form (A) or in a secondhomozygous form (B); and d) selecting at least one pair of doubledhaploid lines that have complementary alleles for at least a subset ofthe genetic markers, wherein each member of the pair is suitable as aparental line for the hybrid organism.

In another aspect, the method for producing parental lines for hybridorganisms, e.g., of pepper variety NUN 70064 PPH, which when crossedreconstitute the genome of pepper variety NUN 70064 PPH, comprising:

-   -   a. defining a set genetic markers that are present a        heterozygous form (H) in a partially heterozygous starting        organism;    -   b. producing at least one further generation from the starting        organism by self-pollination (e.g., F2 or F3 generation);    -   c. selecting at least one pair of progeny organisms in which at        least one genetic marker from the set is present in a        complementary homozygous form (B vs. A, or A vs. B); and    -   d. optionally repeating steps b) and c) until at least one pair        of progeny organisms that have complementary alleles for at        least a subset of the genetic markers has been selected as        parental lines for a hybrid.

The disclosure also relates to a method of producing a combination ofparental lines of a plant of variety NUN 70064 PPH, comprising makingdoubled haploid cells from haploid cells from said plant or plant partof that plant; and optionally crossing these parental lines to produceand collect seeds. In another aspect, the disclosure relates to acombination of parental lines produced by this method. In still anotheraspect, the combination of parental lines can be used to produce a seedor plant of variety NUN 70064 PPH, when these parental lines arecrossed. In still another aspect, the disclosure relates to acombination of parental lines from which a seed or plant having allphysiological and/or morphological characteristics of variety NUN 70064PPH, e.g., determined at the 5% significance level for numericalcharacteristics and determined by type or degree for non-numericalcharacteristics for plants grown under the same conditions.

The disclosure also provides a combination of parental lines which, whencrossed, produce a seed or plant having all physiological and/ormorphological characteristics of variety NUN 70064 PPH, but one, two, orthree which are different (when grown under the same environmentalconditions), as well as a seed or plant having all physiological and/ormorphological characteristics of variety NUN 70064 PPH, but one, two orthree which are different (when grown under the same environmentalconditions).

In another aspect, the disclosure provides a method of determining thegenotype of a plant of the disclosure comprising detecting in the genome(e.g., a sample of nucleic acids) of the plant at least a firstpolymorphism or an allele. The skilled person is familiar with manysuitable methods of genotyping, detecting a polymorphism or detecting anallele including SNP (Single Nucleotide Polymorphism) genotyping,restriction fragment length polymorphism identification (RFLP) ofgenomic DNA, random amplified polymorphic detection (RAPD) of genomicDNA, amplified fragment length polymorphism detection (AFLP), polymerasechain reaction (PCR), DNA sequencing, allele specific oligonucleotide(ASO) probes, and hybridization to DNA microarrays or beads.Alternatively, the entire genome could be sequenced. The method may, incertain aspects, comprise detecting a plurality of polymorphisms in thegenome of the plant, for example by obtaining a sample of nucleic acidfrom a plant and detecting in said nucleic acids a plurality ofpolymorphisms. The method may further comprise storing the results ofthe step of detecting the plurality of polymorphisms on a computerreadable medium.

Also provided is a plant part obtainable from variety NUN 70064 PPH, orfrom progeny of said variety or from a plant having all but one, two, orthree physiological and/or morphological characteristics which aredifferent from those of said variety, or from a vegetatively propagatedplant of variety NUN 70064 PPH (or from its progeny or from a planthaving all or all but one, two, or three physiological and/ormorphological characteristics which are different from those of saidvariety): a fruit, a harvested fruit, a part of a fruit, a leaf, a partof a leaf, pollen, an ovule, a cell, a petiole, a shoot or a partthereof, a stem or a part thereof, a root or a part thereof, a root tip,a cutting, a seed, a part of a seed, seed coat, or another maternaltissue which is part of a seed grown on said variety, or a hypocotyl, acotyledon, a scion, a stock, a rootstock, a pistil, an anther, and aflower or a part thereof.

A part of the plant of pepper variety NUN 70064 PPH (or of progeny ofsaid variety or of a plant having all physiological and/or morphologicalcharacteristics but one, two, or three which are different from those ofsaid variety) encompasses any cells, tissues, organs obtainable from theseedlings or plants, such as but not limited to: a pepper fruit or apart thereof, a cutting, a hypocotyl, a cotyledon, seed coat, or pollen.Such plant parts can be stored and/or processed further.

The disclosure further provides for a food or a feed product comprisinga part of pepper variety NUN 70064 PPH, or a part of progeny of saidvariety, or a part of a plant having all but one, two, or threephysiological and/or morphological characteristics of NUN 70064 PPH,comprising one or more of such parts, optionally processed (such ascanned, chopped, cooked, roasted, in a sauce, in a sandwich, pasted,puréed or concentrated, juiced, frozen, dried, pickled, or powdered).Particularly, the plant part is a pepper fruit or part thereof, and/oran extract from a fruit or another plant part described herein. The foodor feed product may be fresh or processed, e.g., dried, grinded,powdered, pickled, chopped, cooked, roasted, in a sauce, in a sandwich,pasted, puréed or concentrated, juiced, pickled, canned, steamed,boiled, fried, blanched and/or frozen, etc.

In another aspect, the disclosure provides for a pepper fruit of varietyNUN 70064 PPH, or a part of a fruit of said variety. The fruit can be inany stage of maturity, for example, immature or mature. In anotheraspect, the disclosure provides for a container comprising or consistingof a plurality of harvested pepper fruits or parts of fruits of saidvariety, or fruits of progeny thereof, or fruits of a derived variety.

Marketable pepper fruits are generally sorted by size and quality afterharvest. Alternatively, the pepper fruits can be sorted by expectedshelf life or pungency.

In another aspect, the plant, plant part, or seed of pepper variety NUN70064 PPH is inside a container, for example, containers such as cans,boxes, crates, bags, cartons, Modified Atmosphere Packaging, films(e.g., biodegradable films), etc. comprising a plant or a part of aplant (fresh and/or processed) or a seed of pepper variety NUN 70064PPH. In a particular aspect, the container comprises a plurality ofseeds, or a plurality of plant parts of pepper variety NUN 70064 PPH.The seed may be disinfected, primed and/or treated with variouscompounds, such as seed coatings or crop protection compounds. The seedproduces a plant of variety NUN 70064 PPH.

Pepper variety NUN 70064 PPH may also be grown for use as rootstocks(stocks) or scions. Typically, different types of peppers are grafted toenhance disease resistance, which is usually conferred by the rootstock,while retaining the horticultural qualities usually conferred by thescion. It is not uncommon for grafting to occur between cultivatedpepper varieties and related pepper species. Methods of grafting andvegetative propagation are well-known in the art.

In another aspect, the disclosure relates to a plant comprising arootstock or scion of pepper variety NUN 70064 PPH.

All documents (e.g., patent publications) are herein incorporated byreference in their entirety, including the following cited references:

-   Naktuinbow, Calibration book Capsicum annuum L., world-wide web at    naktuinbow.nl.-   UPOV, “Guidelines for the Conduct of Tests for Distinctness,    Uniformity and Stability, TG/76/8 (Geneva 2006, revised 2018); world    wide web at upov.int/under edocs/tgdocs/en/tg076.pdf.-   US Department of Agriculture, Agricultural Marketing Service,    “Objective Plant Description of Variety Pepper (Capsicum spp.)”    world wide web at    ams.usda.gov/services/plant-variety-protection/pvpo-c-forms, under    pepper.-   Hartz, T., et. al, “Bell Pepper Production in California,”    University of California Division of Agriculture and Natural    Resources, Vegetable Production Series, Publication 7217, pp. 1-4.-   Ince, A. G., et al., “Genetic Relationship Within and Between    Capsicum Species”, Biochem Genet, 2010, vol. 48, pp. 83-95.-   Kothari, S. L., et al., “Chili Peppers—A review on Tissue Culture    and Transgenesis”, Biotechnology Advances, 2010, vol. 28, pp. 35-48.-   Martin, E., et al., “Identification of Markers Linked to Agronomic    Traits in Globe Artichoke”, Australian Journal of Crop Science, vol.    1, no. 2, pp. 43-46.-   Needleman, S. B., et. al., “A General Method Applicable to the    Search for Similarities in the Amino Acid Sequence of Two Proteins”,    Journal of Molecular Biology, 1970, vol. 48(3), pp. 443-53-   Nikolova, V., et. al., “Diploidization of Cucumber (Cucumis sativus    L.) Haploids by Colchini Treatment”, Acta Societas Botanicorum    Poloniae, 1996, vol. 65, pp. 311-317.-   Rice, P., et al., “EMBOSS: The European Molecular Biology Open    Software Suite”, Trends in Genetics, 2000, vol. 16, Issue 6. pp.    276-277-   Sang-Gu, K., et al., “Callus Growth and Plant Regeneration in    Diverse Cultivars of Cucumber”, Plant Cell, Tissue and Organ    Culture, 1988, vol. 12, pp. 67-74-   Tiwari, A., et al., “Parthenocarpic Potential in Capsicum annum L.    is Enhanced by Carpelloid Structures and Controlled by Single    Recessive Gene”, BMC Plant Biology, 2011, vol. 11, pp. 2-14, DOI:    10.1186/1471-2229-11-143-   Vos, P., et al., AFLP: A New Technique for DNA Fingerprinting 1995,    Nucleic Acids Research, 1995, vol. 23, No. 21, pp. 4407-4414.-   Wijnker, E., et al., Hybrid Recreation by Reverse breeding in    Arabidopsis thaliana, Nature Protocols, 2014, vol. 9, pp. 761-772.    DOI: doi: 10.1038/nprot.2014.049-   U.S. Pat. No. 8,492,619-   US2006/0037100-   US2015/0245570-   US2015/0126380-   https://www.ams.usda.gov/resources/st470-pepper-   https://anrcatalog.ucanr.edu/pdf/7217.pdf-   http://www.upov.int/edocs/tgdocs/en/tg076_08.pdf-   https://www.wifss.ucdavis.edu/wp-content/uploads/2016/10/Peppers_PDF.pdf

Development of Pepper Variety NUN 70064 PPH

The hybrid NUN 70064 PPH was developed from a male and femaleproprietary inbred line of Nunhems. The female and male parents werecrossed to produce hybrid (F1) seeds of pepper variety NUN 70064 PPH.The seeds of pepper variety NUN 70064 PPH can be grown to produce hybridplants and parts thereof (e.g., pepper fruit). The hybrid NUN 70064 PPHcan be propagated by seeds or vegetatively.

The hybrid variety is uniform and genetically stable. This has beenestablished through evaluation of horticultural characteristics. Severalhybrid seed production events resulted in no observable deviation ingenetic stability. Coupled with the confirmation of genetic stability ofthe female and male parents the Applicant has concluded that peppervariety NUN 70064 PPH is uniform and stable.

Deposit Information

A total of 625 seeds of the hybrid variety NUN 70064 PPH was made andaccepted according to the Budapest Treaty by Nunhems B.V. on ______ atthe NCIMB Ltd., Ferguson Building, Craibstone Estate, Bucksburn,Aberdeen AB21 9YA, United Kingdom (NCIMB). The deposit is assignedAccession Number NCIMB ______. A statement indicating the viability ofthe sample has been provided. A deposit of pepper variety NUN 70064 PPHand of the male and female parent line is also maintained at NunhemsB.V.

Access to the deposits will be available during the pendency of thisapplication to persons determined by the Director of the U.S. PatentOffice to be entitled thereto upon request. Subject to 37 C.F.R. §1.808(b), all restrictions imposed by the depositor on the availabilityto the public of the deposited material will be irrevocably removed uponthe granting of the patent. The deposit will be maintained for a periodof 30 years, or 5 years after the most recent request, or for theenforceable life of the patent whichever is longer and will be replacedif it ever becomes nonviable during that period. Applicant does notwaive any rights granted under this patent on this application or underthe Plant Variety Protection Act (7 U.S.C. § 2321 et seq.).

Characteristics of Pepper Variety NUN 70064 PPH

The most similar variety to variety NUN 70064 PPH is referred to asMixteco, a commercial variety from HM Clause.

In Table 1, the characteristics of pepper variety NUN 70064 PPH and theReference Variety are shown based on a trial under open field conditionsin Los Mochis, Northern Sinaloa, Mexico during the trial season August2021 to April 2022.

In one aspect, the disclosure provides a plant having the physiologicaland morphological characteristics of pepper variety NUN 70064 PPH aspresented in Table 1 when grown under the same environmental conditions,and wherein a representative sample of seed of said pepper variety hasbeen deposited under Accession Number NCIMB ______.

TABLE 1 Characteristics of Pepper Variety NUN 70064 PPH ApplicationReference Variety Variety Characteristics (NUN 70064 PPH) (MIXTECO) Seedcolor: Yellow Species: C. annuum C. annuum C. annuum C. frutescens C.baccatum C. chinense Other Plant: Habit: upright/semi- UprightSemi-spreading spreading/spreading/other Attitude: erect/semi- ErectSemi-erect erect/prostrate/other Basal branches: none/few/many None NoneBranch flexibility: willowy/rigid Rigid Rigid Stem strength: StrongStrong weak/intermediate/strong Leaves: Mature leaf shape: LanceolateLanceolate lanceolate/elliptic Leaf color: light green/medium Dark greenDark green green/dark green/purple/other Leaf and stem pubescence:Absent Absent absent/light/moderate/heavy Margin undulation: absent/veryAbsent absent weak/weak/medium/strong/very strong Blistering:absent/very Absent absent weak/weak/medium/strong/very strong Fruit:Fruit group: Jalapeño Jalapeño Bell/Pimiento/Ancho/Cayenne/Cuban/Jalapeno/Small Hot/Cherry/Short Wax/Long Wax/Tabasco/HabaneroImmature fruit color: light Dark green Dark green green/mediumgreen/dark green/very dark green/yellow/purple/ivory/other Mature fruitcolor: Red Red red/orange/orange- yellow/brown/ivory/green/salmon/ lemonyellow/other Capsaicin in placenta: Present Present absent/presentPungency: sweet/hot Hot Hot Fruit flavor: mild pepper Other Otherflavor/moderate pepper flavor/strong pepper flavor/other Fruitglossiness: Shiny Shiny dull/moderate/shiny Surface smoothness: SmoothSmooth smooth/rough Fruit position: Pendant Pendantupright/horizontal/pendant Calyx shape: cup-shaped/saucer- Saucer-shapeSaucer-shape shaped Fruit base shape: cupped/rounded Rounded RoundedFruit apex shape: pointed, blunt Pointed Pointed Fruit shape: ElongateElongated bell/conical/elongate/oblong/oblate/ globe/other Fruit shapelongitudinal section: Trapezoid Trapezoid flattened/round/heart-shaped/square/rectangular/ trapezoid/narrow triangular/triangular/horn-shaped Fruit shape cross section: Circular Circularelliptic/triangular/quadrangular/ circular Fruit set:scattered/concentrated Scattered Concentrated Interloculary grooves:absent/very Absent Absent shallow/shallow/medium/deep/very deep Numberof locules: Predominantly Predominantly predominantly two/equally twothree three and three/predominantly three/equally three andfour/predominantly four and more Pedicel shape: straight/curved StraightStraight Pedicel cavity: absent/present Absent Absent Maturity: Time ofmaturity: Medium Medium Very early/early/medium/late/ very late Diseaseresistances: Tomabovirus (Tobacco Mosaic Resistant Resistant Virus)Pathotype 0: Tomato Spotted Wilt Virus Susceptible Susceptible (TSWV)Xanthomonas campestris pv. Resistant Resistant Vesicatoria, Pathotype 1

1. A plant, plant part, or seed of pepper variety NUN 70064 PPH, whereina representative sample of seed of said pepper variety is depositedunder Accession Number NCIMB ______.
 2. A plant part of the plant ofclaim 1, wherein said plant part is a fruit, a leaf, pollen, an ovule, acell, a scion, a root, a rootstock, a cutting, or a flower.
 3. A seedthat produces the plant of claim
 1. 4. A pepper plant having all thephysiological and morphological characteristics of the plant of claim 1when grown under the same environmental conditions.
 5. A tissue cultureor cell culture of regenerable cells of the plant or plant part ofclaim
 1. 6. The tissue or cell culture according to claim 5, comprisingcells or protoplasts obtained from a plant part suitable for vegetativereproduction, wherein the plant part is a meristem, a cotyledon, ahypocotyl, a pollen, a leaf, an anther, a root, a root tip, a pistil, apetiole, a flower, a fruit, a seed, a stem, or a stalk.
 7. A method ofproducing the plant of claim 1, or a part thereof, said methodcomprising vegetative propagation of at least a part of pepper varietyNUN 70064 PPH, wherein a representative sample of seed of said peppervariety is deposited under Accession Number NCIMB ______.
 8. The methodof claim 7, wherein said part is a cutting, a cell culture, or a tissueculture.
 9. A vegetatively propagated plant, or part thereof, producedby the method of claim 7, wherein the plant, or part thereof has all ofthe physiological and morphological characteristics of the plant ofvariety NUN 70064 PPH when grown under the same environmentalconditions, wherein a representative sample of seed of said peppervariety is deposited under Accession Number NCIMB ______.
 10. A methodof producing a pepper plant, said method comprising crossing the plantof claim 1 with itself or a second pepper plant, selecting a progenypepper plant from said crossing, allowing the progeny pepper plant toform seed, wherein a representative sample of seed of said peppervariety NUN 70064 PPH is deposited under Accession Number NCIMB ______.11. A method of introducing a desired trait into the plant of claim 1,comprising transforming the plant of pepper variety NUN 70064 PPH with atransgene that confers the desired trait, wherein the plant comprisesthe desired trait and otherwise has all of the physiological andmorphological characteristics of the plant of pepper variety NUN 70064PPH.
 12. A pepper plant produced by the method of claim 11, wherein thetransformed plant otherwise comprises the desired trait and otherwisehas all the physiological and morphological characteristics of peppervariety NUN 70064 PPH.
 13. A method of making doubled haploid cells ofthe plant of claim 1, said method comprising making double haploid cellsfrom haploid cells of the plant or seed of pepper variety NUN 70064 PPH,wherein a representative sample of seed of said pepper variety isdeposited under Accession Number NCIMB ______.
 14. A plant comprisingthe scion or rootstock of claim
 2. 15. A container comprising the plant,plant part, or seed of claim
 1. 16. A food or feed product or aprocessed product comprising the plant part of claim 2, wherein saidplant part comprises at least a regenerable cell of pepper variety NUN70064 PPH.
 17. A method of introducing a single locus conversion intothe plant of claim 1 comprising: a. crossing the plant of claim 1 with asecond pepper plant comprising a desired single locus conversion toproduce F1 progeny plants; b. selecting F1 progeny plants that have thesingle locus conversion to produce selected F1 progeny plants; c.crossing the selected F1 progeny plants with pepper variety NUN 70064PPH to produce backcross progeny plants; d. selecting backcross progenyplants that have the single locus conversion and otherwise comprise allof the physiological and morphological characteristics of pepper varietyNUN 70064 PPH to produce selected backcross progeny plants; and e.repeating steps (c) and (d) one or more times in succession to produceselected second or higher backcross progeny plants that comprise thesingle locus conversion and otherwise comprise all of the physiologicaland morphological characteristics of the plant of pepper variety NUN70064 PPH, wherein a representative sample of seed of said peppervariety is deposited under Accession Number NCIMB ______.
 18. A pepperplant produced by the method of claim 17, wherein the plant comprisesthe single locus conversion and otherwise has all the physiological andmorphological characteristics of pepper variety NUN 70064 PPH, andwherein the single locus confers male sterility, herbicide tolerance,insect resistance, pest resistance, disease resistance, environmentalstress tolerance, modified carbohydrate metabolism, or modified proteinmetabolism.
 19. A method of producing a modified pepper plant, saidmethod comprising mutating a target gene by targeted gene editing in apepper plant or plant part of pepper variety NUN 70064 PPH, wherein arepresentative sample of seed of said pepper variety is deposited underAccession Number NCIMB ______, wherein the target gene modifies adesired trait and wherein the desired trait is male sterility, herbicidetolerance, insect resistance, pest resistance, disease resistance,environmental stress tolerance, modified carbohydrate metabolism, ormodified protein metabolism.
 20. A pepper plant produced by the methodof claim
 19. 21. A method for determining the genotype of the plant ofclaim 1, said method comprising obtaining a sample of nucleic acids fromsaid plant and detecting in said nucleic acids a plurality ofpolymorphisms, thereby determining the genotype of the plant, andstoring the results of detecting the plurality of polymorphisms on acomputer readable medium.
 22. A method of producing a pepper fruit, saidmethod comprising growing the plant of claim 1 until it develops atleast a leaf or head, and collecting the leaf or head.
 23. The fruitproduced by the method of claim
 22. 24. A container comprising the fruitor part thereof produced by the method of claim
 21. 25. A method fordeveloping a pepper plant in a pepper breeding program, said methodcomprising applying plant breeding techniques comprising recurrentselection, backcrossing, pedigree breeding, mass selection, mutationbreeding, genetic marker enhanced selection, or genetic transformationto the plant of claim 1 or part thereof, wherein said plant breedingtechniques result in a development of a pepper plant.
 26. A method ofproducing a pepper plant derived from the plant of claim 1 comprising:a. preparing a progeny pepper plant derived from pepper variety NUN70064 PPH by crossing the plant of pepper variety NUN 70064 PPH withitself or with a second pepper plant, wherein a representative sample ofseed of said pepper variety has been deposited under Accession NumberNCIMB ______; b. crossing the progeny plant with itself or a secondpepper plant to produce seed of a progeny plant of the subsequentgeneration; c. growing a progeny plant of the subsequent generation fromsaid seed and crossing the progeny plant of the subsequent generationwith itself or a second pepper plant; and d. repeating step (b) and (c)for at least one more generation to produce a pepper plant derived frompepper variety NUN 70064 PPH.
 27. A method of producing a pepper seed,said method comprising crossing pepper plants and harvesting theresultant seed, wherein at least one pepper plant is the plant of claim1, wherein a representative sample of seed of said pepper variety NUN70064 PPH is deposited under Accession Number NCIMB ______.